Vaccine for control of beet curly top virus infection of plants

Abstract

The disclosure provides plant vaccine compositions and methods for inducing immunity to Beet Curly Top Virus (BCTV) and related curtoviruses in a susceptible plant.

What is claimed is: 
     
       1. A plant vaccine composition for inducing immunity to Beet Curly Top Virus (BCTV) in a susceptible plant, the plant vaccine comprising: an expression vector comprising a heterologous promoter sequence operably linked to an isolated BCTV vaccine nucleic acid sequence that is selected from the group consisting of (i) SEQ ID NO:14 and (ii) both SEQ ID NO:10 and SEQ ID NO:11. 
     
     
       2. A plant vaccine composition for inducing immunity to Beet Curly Top Virus (BCTV) in a susceptible plant, the plant vaccine comprising: an expression vector comprising a heterologous promoter operably linked to SEQ ID NO:10 wherein SEQ ID NO:10 is operably linked to an intron, and the intron is operably linked to SEQ ID NO:11. 
     
     
       3. The plant vaccine composition of  claim 1 , wherein the vaccine induces immunity to Beet Curly Top Viruses that are members selected from the group consisting of Beet Severe Curly Top Virus (BSCTV) and Beet Mild Curly Top Virus (BMCTV). 
     
     
       4. The plant vaccine composition of  claim 1 , wherein the susceptible plant is a member selected from the group consisting of sugar beet ( Beta vulgaris ), tomato ( Solanum lycopersicum ), bean ( Phaseolus vulgaris ), and pepper ( Capsicum  spp.). 
     
     
       5. The plant vaccine composition of  claim 4 , wherein the susceptible plant is sugar beet. 
     
     
       6. The plant vaccine composition of  claim 4 , wherein the susceptible plant is tomato. 
     
     
       7. A method for inducing immunity to Beet Curly Top Virus (BCTV) in a susceptible plant, thereby protecting the susceptible plant from infection with BCTV, the method comprising:
 (i) transfecting the susceptible plant with a suspension of  Agrobacterium tumefaciens , wherein the  Agrobacterium tumefaciens  comprises an expression vector comprising a heterologous promoter operably linked to an isolated BCTV vaccine nucleic acid sequence that is selected from the group consisting of (i) SEQ ID NO:14 and (ii) the combination of SEQ ID NO:10 and SEQ ID NO:11, to generate a transformed plant; and 
 (ii) selecting said transformed plant that expresses said isolated BCTV vaccine nucleic acid sequence to produce a vaccinated plant; 
 wherein expression of the isolated BCTV vaccine nucleic acid sequence in the vaccinated plant induces silencing of the BCTV, thereby protecting the susceptible plant from infection with BCTV. 
 
     
     
       8. A method for inducing immunity to Beet Curly Top Virus (BCTV) in a susceptible plant, thereby protecting the susceptible plant from infection with BCTV, the method comprising:
 (i) transfecting the susceptible plant with a suspension of  Agrobacterium tumefaciens , wherein the  Agrobacterium tumefaciens  comprises an expression vector comprising a heterologous promoter operably linked to SEQ ID NO:10 wherein SEQ ID NO:10 is operably linked to an intron, and the intron is operably linked to SEQ ID NO:11 to generate a transformed plant, 
 (ii) selecting said transformed plant that expresses said SEQ ID NO:10, intron and SEQ ID NO:11 to produce a vaccinated plant; 
 wherein expression of the expression vector in the vaccinated plant induces silencing of the BCTV thereby protecting the susceptible plant from infection with BCTV. 
 
     
     
       9. The method of  claim 7 , wherein the transfection is transient transfection. 
     
     
       10. The method of  claim 9 , wherein transient transfection is carried out by direct injection of the suspension of  Agrobacterium tumefaciens  into a stem of the susceptible plant. 
     
     
       11. The method of  claim 9 , wherein transient transfection comprises spraying the suspension of  Agrobacterium tumefaciens  onto one or more leaves of the susceptible plant. 
     
     
       12. The method of  claim 7 , wherein the method induces immunity to Beet Curly Top Viruses that are members selected from the group consisting of Beet Severe Curly Top Virus (BSCTV) and Beet Mild Curly Top Virus (BMCTV). 
     
     
       13. The method of  claim 7 , wherein the susceptible plant is a member selected from the group consisting of sugar beet ( Beta vulgaris ), tomato ( Solanum lycopersicum ), bean ( Phaseolus vulgaris ), and pepper ( Capsicum  spp.). 
     
     
       14. The method of  claim 13 , wherein the susceptible plant is sugar beet. 
     
     
       15. The method of  claim 13 , wherein the susceptible plant is tomato. 
     
     
       16. A transformed plant comprising an expression vector wherein said expression vector comprises a heterologous promoter operably linked to a nucleic acid selected from the group consisting of (i) SEQ ID NO:14 and (ii) both SEQ ID NO:10 and SEQ ID NO:11. 
     
     
       17. The transformed plant of  claim 16 , wherein said plant is selected from the group consisting of sugar beet ( Beta vulgaris ), tomato ( Solanum lycopersicum ), bean ( Phaseolus vulgaris ), and pepper ( Capsicum  spp.). 
     
     
       18. The method of  claim 8 , wherein the transfection is transient transfection. 
     
     
       19. The method of  claim 18 , wherein transient transfection is carried out by direct injection of the suspension of  Agrobacterium tumefaciens  into a stem of the susceptible plant. 
     
     
       20. The method of  claim 18 , wherein transient transfection comprises spraying the suspension of  Agrobacterium tumefaciens  onto one or more leaves of the susceptible plant. 
     
     
       21. The method of  claim 18  wherein said transient transfection comprises dipping said susceptible plant into said suspension of  Agrobacterium tumefaciens.    
     
     
       22. The method of  claim 9  wherein said transient transfection comprises dipping said susceptible plant into said suspension of  Agrobacterium tumefaciens.    
     
     
       23. The plant vaccine composition of  claim 1  further comprising a carrier. 
     
     
       24. The plant vaccine composition of  claim 23 , wherein said carrier comprises 2-(4-morpholine)-ethane sulfonic acid (MES), MgCl 2  and acetosyringone. 
     
     
       25. The plant vaccine composition of  claim 2  further comprising a carrier. 
     
     
       26. The plant vaccine composition of  claim 25 , wherein said carrier comprises MES, MgCl 2  and acetosyringone. 
     
     
       27. The method of  claim 7  wherein said suspension of  Agrobacterium tumefaciens  further comprises MES, MgCl 2  and acetosyringone. 
     
     
       28. The method of  claim 8  wherein said suspension of  Agrobacterium tumefaciens  further comprises MES, MgCl 2  and acetosyringone.